Commercial feasibility and advantages of micro propagation

Commercial feasibility and advantages of micro propagation:-

Commercial Feasibility:-

1. Market Scenario:-

> Demand for tissue cultured plantlets is growing rapidly. India, with its low cost skilled labour as well as scientific manpower (both of which are essential for tissue culture) has a natural advantage. Additional favourable factors are the wide range of plant biodiversity in the country and favorable tropical climate (which enables greenhouses with low energy consumption).

> The potential for the domestic market is enormous and by conservative estimates it is around Rs. 200 crores with an annual growth rate of 20%. There are more than 70 established commercial tissue culture units. Their production capacity ranges between 0.5 million to 10 million plants per annum with an aggregate production capacity of about 200 million plantlets per year.

> The protocols have either been developed in-house or transferred through the various research institutions and universities engaged in development of the protocols through support of the Department of Biotechnology (DBT) Currently, the focus of the companies is mainly banana, floriculture,

sugarcane and potato.

> With increasing awareness about the advantages of tissue culture raised plants in improving yield and quality, their domestic consumption is also increasing optimistically. The major consumers of tissue culture raised plants are the State Agriculture Department, Agri Export Zones (AEZs), State agencies such as Spice Board, sugar industry and private farmers. The paper industry, medicinal plant industry and State Forest Departments are using tissue culture raised plants in a limited scale. Also a number of progressive farmers and nurseries in the states are the major consumers of Tissue culture plants particularly for flowers, banana, sugarcane and medicinal plants.

2. Establishment of Commercial Plant Tissue Culture Unit:- Commercial plant tissue culture unit consists of the following components:

a. Storage room for chemicals:- It is advisable to have a separate area for storage of chemicals, apparatus and equipments. Chemicals required in small amounts should not be purchased in large quantities as they may lose their activity, pick up moisture or get contaminated. Such problems can be

overcome by purchasing small lots on a regular basis.

b. Washing and Media Preparation Room:- 

> The glassware washing area should be located near the sterilization room. This area should have at least one large sink but two sinks are preferable with running tap water. Adequate workspace is required on each sides of the sink; this space is used for glassware soaking and drainage. Plastic netting can be placed on surfaces near the sink to reduce glassware breakage and enhance water drainage. The outlet

pipe from the sink should be of PVC to resist damage from acids and alkalis. Both hot and cold water should be available and the water still and de-ionisation unit should be located nearby. The washing room should be swapped periodically. Mobile drying racks can be used and lined with cheesecloth to prevent water dripping and loss of small objects. 

> Ovens or hot air-cabinets should be located close to the glassware washing and storage area. Dust-proof cabinets and storage containers should be installed to allow for easy access to glassware. When culture vessels are removed from the growth area, they are often autoclaved to kill contaminants and to soften semi-solid media. It should be possible to move the vessels easily to the washing area. The glassware storage area should be close to the wash area to expedite storage and access for media preparation.

> The media preparation room should have smooth walls and floors, which enable easy cleaning to maintain a high degree of cleanliness.

> Minimum number of doors and windows should be provided in this room but within the local fire safety regulations. Media preparation area should be equipped with both tap and purified water. An appropriate system for water purification must be selected and fitted after careful consideration of the cost and quality. A number of electrical appliances are required for media preparation; hence, it is essential to have safety devices like fire extinguisher, fire blanket and a first aid kit in the media preparation room. A variety of glassware, plastic ware and stainless steel apparatus is required for measuring, mixing, and media storage. These should be stored in the cabinets built under the worktables and taken out for use as and when required. 

> The water source and glassware storage area should be in or near the media preparation area. The workbench tops should be made with plastic laminate surfaces that can tolerate frequent cleaning. Media storage room should have capacity to storage the media for at least 7 days. Sterility Class

1,00,000 is desirable for media storage room. 

c. Inoculation Room:-

> The most important work area is the Inoculation room where the core activity takes place. The transfer area needs to be as clean as possible with minimal air disturbance. Walls and floors of the Inoculation room must be smooth to ensure frequent cleaning. The doors and windows should be minimal to prevent contamination, but within local safety code. There is no special lighting requirement in the transfer room. The illumination of the laminar airflow chamber is sufficient for work.

> Sterilization of the instruments can be done with glass-bead sterilizers or flaming after dipping in alcohol, usually ethanol. The culture containers should be stacked on mobile carts (trolleys) to facilitate easy movement from the medium storage room to the transfer room, and finally to the culture room. Fire extinguishers and first aid kits should be provided in the transfer room as a safety measure. Special laboratory shoes and coats should be worn in this area. Ultraviolet (UV) lights are sometimes installed in transfer areas to disinfect the room; these lights should be used only when people and plant material are not in the room. Sterility Class 1,00,000 is desirable for inoculation room which can be achieved through installation of pressurized air module or air handling unit.

d. Growth Room:- 

> Culture room is an equally important area where plant cultures are maintained under controlled environmental conditions to achieve optimal growth. It is advisable to have more than one growth room to provide varied culture conditions since different plant species may have different requirements of light and temperature during in vitro culture.

> Also, in the event of the failure of cooling or lighting in one room, the plant cultures can be moved to another room to prevent loss of cultures. In the growth room, the number of doors should be minimal to prevent contamination. The culture containers can be placed on either fixed or mobile shelves. Mobile shelves have the advantage of providing access to cultures from both sides of the shelves. The height of the shelves should not exceed 2m.

> The primary source of illumination in the growth room is normally from the lights mounted on the shelves. Overhead light sources can be minimized, as they would be in use only while working during the dark cycle. Plant cultures may not receive uniform light from the conventional downward illumination. Lights directly fitted to the racks create uneven heat distribution. Sideways illumination is an alternative, which requires less number of lights, and provides more uniform lighting. But care has to be taken not to break the lights while moving the cultures across the shelves. Sterility Class 1,00,000 is desirable for growth room.

Advantages of Micro-propagation:- Micro-propagation has several advantages over conventional methods of propagation such as:

1. Rapid multiplication:- Micro-propagation offers rapid multiplication of desired plant speceis.

2. Requirement of only limited number of explants:-  Small pieces of plant (explants)/tissue can be used to produce a large number of plants in a relatively small space.

3. Uniform or true to type plants:- Micro-propagation provides a high degree of phenotypic/physical uniformity. Since the production cycle takes place under controlled conditions, proper planning and scheduling based on the market demand is possible. The resulting product has very high degree of uniformity compared with traditionally propagated plants.

4. Germplasm storage:- Plants can be stored in vitro in a small space and less labour is required for maintenance of stock plants.

5. Disease free planting material:- Plantlets produced by tissue culture are usually disease free. With proper diagnosis and treatments, elimination of fungus, bacteria and virus prior to large scale propagation is possible. With the help of seroloical and molecular technique it is possible to index virus of mother plant/explant which is to be used for mass multiplication. 

6. Growth manipulation:- Nutrient levels, light, temperature and other factors can be more effectively controlled to manipulate the growth, multiplication and regeneration.

7. Round the year production:- Micro-propagation is independent of season. As micro - propagation could be carried out throughout the year; production cycle can be scheduled to meet peak demands.

8. Rapid propagation is possible:- For species that have long generation time, low levels of seed production, or seeds that do not readily germinate, rapid propagation is possible through tissue culture.

9. No need to wait:- The time required is much shortened, no need to wait for the whole life cycle of seed development.